Junya Li

Investigation on BRCA1 SNPs and its effects on mastitis in Chinese commercial cattle.

The main objective of this study was to investigate whether the bovine breast cancer 1 (BRCA1) gene was associated with mastitis resistance in Chinese commercial cattle. A total of 51 SNPs were screened from public data resources and DNA sequencing. Three SNPs (c.5682G>C,c.26198C>T and c.46126G>T) were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and created restriction site PCR (CRS-PCR) methods and 21 combinations of these SNPs were observed. The single SNP and their genetic effects on somatic cell score (SCS) were evaluated and a significant association with SCS was found in c.46126G>T. The mean SCS of individuals with genotype KK was significantly lower than those of genotypes KL and LL. The results of combined genotypes analysis of three SNPs showed that HHLLNN genotype with the highest SCS was easily for the mastitis susceptibility, whereas GGKKMM genotype with the lowest SCS was favorable for the mastitis resistance. The information provided in the present study will be very useful for improving mastitis resistance in dairy cattle by marker-assisted selection (MAS).

SNPs identification and its correlation analysis with milk somatic cell score in bovine MBL1 gene

The objective of this study was to investigate the single nucleotide polymorphisms (SNPs) within bovine binding lectin-liver (A) gene (MBL1) and to explore its correlation analysis with milk somatic cell score (SCS) which reflects mastitis resistance in cattle. Through polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), created restriction site-PCR (CRS-PCR) and DNA sequencing methods, three allelic variant corresponding to the G→A mutation at c.1252G>A in intron1, G→A mutation at c.2534G>A and T→C mutation at c.2569T>C in exon2 of bovine MBL1 gene, could be detected, respectively. The c.2534G>A was a nonsynonymous mutation, resulting in Valine (Val) to Isoleucine (Ile) amino acid replacement (p.Val24Ile). The correlation analysis between the MBL1 SNPs gene and milk SCS were analyzed and a significant correlation with milk SCS was detected in c.2534G>A. The value of milk SCS for individuals with genotype GG was significantly lower than those of genotype GA and AA. Results showed that genotype GG with the lowest milk SCS was favorable for mastitis resistance, whereas genotype AA with the highest milk SCS was easily for mastitis susceptibility. Although more investigations are needed to better clarify the role of these SNPs on mastitis resistance, MBL1 polymorphism appears to be a promising indirect marker to improve dairy mastitis resistance traits in cattle.

Effects of DGAT1 gene on meat and carcass fatness quality in Chinese commercial cattle

This study was designed to investigate the candidate single nucleotide polymorphisms (SNPs) in the exon’s region of bovine diacylglycerol O-acyltransferase (DGAT1) gene using bioinformatics and experimental methods. A total of 17 SNPs were screened from public data resources and DNA sequencing. Three SNPs (c.572A>G, c.1241C>T and c.1416T>G) of these candidate SNPs were genotyped by created restriction site-polymerase chain reaction (CRS-PCR) methods. The gene-specific SNP markers and their effects on meat and carcass fatness quality traits were evaluated in Chinese commercial cattle. The c.572A>G and c.1416T>G significantly effected on backfat thickness, longissimus muscle area, marbling score, fat color and Warner-Bratzler shear force. No significant association was detected between the c.1241C>T and measured traits. Results from this study suggested that the SNP markers may be effective for the marker-assisted selection of meat and carcass fatness quality traits, and added new evidence that DGAT1 gene is an important candidate gene for the improvement of meat and carcass fatness quality in beef cattle industry.

Association analysis of thyroglobulin gene variants with carcass and meat quality traits in beef cattle.

Thyroid hormones play an important role in regulating the metabolism and can affect the homeostasis of fat deposition. The gene encoding thyroglobulin (TG), producing the precursor for thyroid hormones, has been proposed as a positional and functional candidate gene for a QTL with an effect on fat deposition. In the present study, we identified 6 novel SNPs at the 3’ flanking region of theTG gene. The SNP marker association analysis indicated that the SNP markers G133C, G156A, C220T and A506C were significantly associated with marbling score (P<0.05,N=271). Animals with the new homozygote genotype had higher marbling scores than those with the other genotypes. Besides, the linkage disequilibrium analysis indicated that these 4 SNPs were completely linked (r2 = 1). Results of this study suggest that theTG-gene-specific SNP may be a useful marker for meat quality traits in future marker-assisted selection programmes in beef cattle.

BRCA1: a new candidate gene for bovine mastitis and its association analysis between single nucleotide polymorphisms and milk somatic cell score

Bovine mastitis is a very complex and common disease of dairy cattle and a major source of economic losses to the dairy industry worldwide. In this study, the bovine breast cancer 1, early onset gene (BRCA1) was taken as a candidate gene for mastitis resistance. The main object of this study was to investigate whether the BRCA1 gene was associated with mastitis in cattle. Through DNA sequencing, Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and Created Restriction Site PCR (CRS-PCR) methods, three SNPs (G22231T, T25025A, and C28300A) were detected and twenty-four combinations of these SNPs were observed. The single SNP and their genetic effects on somatic cell score (SCS) were evaluated and a significant association with SCS was found in C28300A. The mean of genotype EE was significantly lower than those of genotypes EF and FF. The results of combined genotypes analysis of three SNPs showed that BBDDFF genotype with the highest SCS were easily for the mastitis susceptibility, whereas AACCEE genotype with the lowest SCS were favorable for the mastitis resistance. The information provided in the present study will be very useful for improving mastitis resistance in dairy cattle by marker-assisted selection.

A genome‐wide scan for copy number variations using high‐density single nucleotide polymorphism array in Simmental cattle

Copy number variations (CNVs) have recently been identified as promising sources of genetic variation, complementary to single nucleotide polymorphisms (SNPs). As a result, detection of CNVs has attracted a great deal of attention. In this study, we performed genome-wide CNV detection using Illumina Bovine HD BeadChip (770k) data on 792 Simmental cattle. A total of 263 CNV regions (CNVRs) were identified, which included 137 losses, 102 gains and 24 regions classified as both loss and gain, covering 35.48 Mb (1.41%) of the bovine genome. The length of these CNVRs ranged from 10.18 kb to 1.76 Mb, with an average length of 134.78 kb and a median length of 61.95 kb. In 136 of these regions, a total of 313 genes were identified related to biological functions such as transmembrane activity and olfactory transduction activity. To validate the results, we performed quantitative PCR to detect nine randomly selected CNVRs and successfully confirmed seven (77.6%) of them. Our results present a map of cattle CNVs derived from high-density SNP data, which expands the current CNV map of the cattle genome and provides useful information for investigation of genomic structural variation in cattle.

Genome-wide association studies using haplotypes and individual SNPs in Simmental cattle.

Recent advances in high-throughput genotyping technologies have provided the opportunity to map genes using associations between complex traits and markers. Genome-wide association studies (GWAS) based on either a single marker or haplotype have identified genetic variants and underlying genetic mechanisms of quantitative traits. Prompted by the achievements of studies examining economic traits in cattle and to verify the consistency of these two methods using real data, the current study was conducted to construct the haplotype structure in the bovine genome and to detect relevant genes genuinely affecting a carcass trait and a meat quality trait. Using the Illumina BovineHD BeadChip, 942 young bulls with genotyping data were introduced as a reference population to identify the genes in the beef cattle genome significantly associated with foreshank weight and triglyceride levels. In total, 92,553 haplotype blocks were detected in the genome. The regions of high linkage disequilibrium extended up to approximately 200 kb, and the size of haplotype blocks ranged from 22 bp to 199,266 bp. Additionally, the individual SNP analysis and the haplotype-based analysis detected similar regions and common SNPs for these two representative traits. A total of 12 and 7 SNPs in the bovine genome were significantly associated with foreshank weight and triglyceride levels, respectively. By comparison, 4 and 5 haplotype blocks containing the majority of significant SNPs were strongly associated with foreshank weight and triglyceride levels, respectively. In addition, 36 SNPs with high linkage disequilibrium were detected in the GNAQ gene, a potential hotspot that may play a crucial role for regulating carcass trait components.

Association of the leptin gene E2-169T>C and E3-299T>A mutations with carcass and meat quality traits of the Chinese Simmental-cross steers.

Leptin is a hormone affecting the regulation of body composition, energy balance, and meat quality in mammals. The objective of this study was to evaluate the association of novel single nucleotide polymorphisms in coding region for leptin gene with carcass and meat quality traits of Chinese Simmental-cross steers. Two SNPs (E2-169 T>C and E3-299 T>A) were genotyped on 135 crossbred bulls. The 45 traits being measured included dressing percentage, dressed weight, marbling score, muscle color score, backfat thickness, fatty acid content, etc. Statistical analysis revealed that two SNPs in the exon of leptin gene were associated with the carcass and meat quality traits. The C-bearing genotypes (CC or TC) of E2-169 T>C (C57R) showed higher dressed weight, thickness of loin, MCS, FCS, intramuscular fat content, and polyunsaturated fatty acid content (P<0.05). E3-299 >A(S100T) also showed a significant association with the carcass traits (dressing percentage, living QIB) and fatty acid content in Simmental-cross steers(P<0.05). Our findings suggested that polymorphisms in leptin might be one of the important genetic factors that influence carcass yield and meat quality in beef cattle, and it may be a useful marker for meat quality traits in future marker-assisted selection programs in beef cattle breeding and production.

Effect of culture conditions on the plant regeneration via organogenesis from cotyledonary node of cowpea ( Vigna unguiculata L. Walp)

A rapid and efficient regeneration system via organogenesis from cotyledonary node explants of cowpea (Vigna unguiculata L. Walp) has been established. The cotyledonary node explants excised from 4 days old seedlings, placed in vitro on medium containing salts of Murashige and Skoog and vitamins of Gamborg’s media (MSB 5 ). Adventitious shoots occurred at the basal end of the initiated axillary buds that preexisted at the node regions. BAP at 1.25 mg/l was the optimum for shoot induction. The combination of BAP with IBA had worthless effect on shoots proliferation. The number of adventitious buds was promoted when the seeds were preconditioned with appropriate concentrations of BAP (2 to 3 mg/l), whereas was depressed with higher concentrations of BAP (5 to 15 mg/l). The regeneration system was further optimized due to the presence of cotyledons attaching to the cotyledonary node explants. Explants with two entire cotyledons from 4 days old seedlings produced the greater number of shoots (7.83) after 3 weeks on MSB5 medium supplemented with 1.25 mg/l BAP. Regenerated shoots could well elongate on regulator-free basal medium and well root with 100% of success on the half strength medium supplemented with various concentrations of IBA (0, 0.1, 0.3, and 0.5 mg/l). The regenerated plantlets were cultured on the pots containing sterilized vermiculite and soil (1:1) with 27% of survival. Key words : Cowpea, plant regeneration, cotyledonary node, organogenesis.

Pathway-Based Genome-Wide Association Studies for Two Meat Production Traits in Simmental Cattle

Most single nucleotide polymorphisms (SNPs) detected by genome-wide association studies (GWAS), explain only a small fraction of phenotypic variation. Pathway-based GWAS were proposed to improve the proportion of genes for some human complex traits that could be explained by enriching a mass of SNPs within genetic groups. However, few attempts have been made to describe the quantitative traits in domestic animals. In this study, we used a dataset with approximately 7,700,000 SNPs from 807 Simmental cattle and analyzed live weight and longissimus muscle area using a modified pathway-based GWAS method to orthogonalise the highly linked SNPs within each gene using principal component analysis (PCA). As a result, of the 262 biological pathways of cattle collected from the KEGG database, the gamma aminobutyric acid (GABA)ergic synapse pathway and the non-alcoholic fatty liver disease (NAFLD) pathway were significantly associated with the two traits analyzed. The GABAergic synapse pathway was biologically applicable to the traits analyzed because of its roles in feed intake and weight gain. The proposed method had high statistical power and a low false discovery rate, compared to those of the smallest P-value and SNP set enrichment analysis methods.